JPH01246299A - Ergosterol derivative and production thereof - Google Patents
Ergosterol derivative and production thereofInfo
- Publication number
- JPH01246299A JPH01246299A JP7187988A JP7187988A JPH01246299A JP H01246299 A JPH01246299 A JP H01246299A JP 7187988 A JP7187988 A JP 7187988A JP 7187988 A JP7187988 A JP 7187988A JP H01246299 A JPH01246299 A JP H01246299A
- Authority
- JP
- Japan
- Prior art keywords
- formula
- extract
- diene
- 5alpha
- 3beta
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000004519 manufacturing process Methods 0.000 title claims description 4
- 150000002137 ergosterols Chemical class 0.000 title description 16
- 150000001875 compounds Chemical class 0.000 claims abstract description 12
- NYMHFYDQBMRBMB-UHFFFAOYSA-N 9-hydroxycerevisterol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)C=CC(C)C(C)C)CCC33)C)(O)C3=CC(O)C21O NYMHFYDQBMRBMB-UHFFFAOYSA-N 0.000 claims abstract 2
- 239000000126 substance Substances 0.000 claims description 10
- GQVCGTRDXSDAHC-FTMBUFPESA-N (3s,5r,6r,9s,10r,13r,14r,17r)-17-[(2r,5r)-5,6-dimethylhept-3-en-2-yl]-6-methoxy-10,13-dimethyl-1,2,3,4,6,9,11,12,14,15,16,17-dodecahydrocyclopenta[a]phenanthrene-3,5-diol Chemical group C([C@@]12C)C[C@H](O)C[C@]1(O)[C@H](OC)C=C1[C@@H]2CC[C@]2(C)[C@@H]([C@H](C)C=C[C@H](C)C(C)C)CC[C@H]21 GQVCGTRDXSDAHC-FTMBUFPESA-N 0.000 claims description 4
- GQVCGTRDXSDAHC-UHFFFAOYSA-N (22E,24R)-24-methyl-6beta-methoxy-5alpha-cholesta-7,22-diene-3beta,5-diol Natural products CC12CCC(O)CC1(O)C(OC)C=C1C2CCC2(C)C(C(C)C=CC(C)C(C)C)CCC21 GQVCGTRDXSDAHC-UHFFFAOYSA-N 0.000 claims description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims 1
- 125000000468 ketone group Chemical group 0.000 claims 1
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 abstract description 27
- 238000000605 extraction Methods 0.000 abstract description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 abstract description 6
- 238000000622 liquid--liquid extraction Methods 0.000 abstract description 4
- 206010028980 Neoplasm Diseases 0.000 abstract description 3
- 201000011510 cancer Diseases 0.000 abstract description 3
- 238000004809 thin layer chromatography Methods 0.000 abstract description 3
- 239000002253 acid Substances 0.000 abstract description 2
- 238000004440 column chromatography Methods 0.000 abstract description 2
- 239000012141 concentrate Substances 0.000 abstract description 2
- GUERPVMWCQXYEU-ZFZPKPCHSA-N 3beta,5alpha,9alpha-trihydroxyergosta-7,22-dien-6-one Natural products C1[C@@H](O)CC[C@]2(C)[C@](CC[C@@]3([C@@H]([C@H](C)C=C[C@H](C)C(C)C)CC[C@H]33)C)(O)C3=CC(=O)[C@]21O GUERPVMWCQXYEU-ZFZPKPCHSA-N 0.000 abstract 2
- -1 3beta,5alpha,6beta,9alpha-tetrahydroxyergosta-7,22-diene Chemical compound 0.000 abstract 1
- 241001327634 Agaricus blazei Species 0.000 abstract 1
- 239000003513 alkali Substances 0.000 abstract 1
- 235000013399 edible fruits Nutrition 0.000 abstract 1
- 239000000706 filtrate Substances 0.000 abstract 1
- 239000000463 material Substances 0.000 abstract 1
- 238000002360 preparation method Methods 0.000 abstract 1
- 238000000638 solvent extraction Methods 0.000 abstract 1
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 21
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 12
- 210000004027 cell Anatomy 0.000 description 8
- 230000022534 cell killing Effects 0.000 description 7
- 239000003960 organic solvent Substances 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 235000001674 Agaricus brunnescens Nutrition 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000002378 acidificating effect Effects 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 230000007935 neutral effect Effects 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 241000222518 Agaricus Species 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 206010008342 Cervix carcinoma Diseases 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 2
- 238000005377 adsorption chromatography Methods 0.000 description 2
- 201000010881 cervical cancer Diseases 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 230000000445 cytocidal effect Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 description 2
- OILXMJHPFNGGTO-UHFFFAOYSA-N (22E)-(24xi)-24-methylcholesta-5,22-dien-3beta-ol Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)C=CC(C)C(C)C)C1(C)CC2 OILXMJHPFNGGTO-UHFFFAOYSA-N 0.000 description 1
- RQOCXCFLRBRBCS-UHFFFAOYSA-N (22E)-cholesta-5,7,22-trien-3beta-ol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)C=CCC(C)C)CCC33)C)C3=CC=C21 RQOCXCFLRBRBCS-UHFFFAOYSA-N 0.000 description 1
- ZHBXGHWSDHVEKS-SHMGQAMKSA-N (3S,5R,6R,9R,10R,13R,14R,17R)-17-[(2R,5R)-5,6-dimethylhept-3-en-2-yl]-6-methoxy-10,13-dimethyl-2,3,4,6,11,12,14,15,16,17-decahydro-1H-cyclopenta[a]phenanthrene-3,5,9-triol Chemical compound CO[C@@H]1C=C2[C@@H]3CC[C@H]([C@H](C)C=C[C@H](C)C(C)C)[C@@]3(C)CC[C@]2(O)[C@@]2(C)CC[C@H](O)C[C@]12O ZHBXGHWSDHVEKS-SHMGQAMKSA-N 0.000 description 1
- OQMZNAMGEHIHNN-UHFFFAOYSA-N 7-Dehydrostigmasterol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)C=CC(CC)C(C)C)CCC33)C)C3=CC=C21 OQMZNAMGEHIHNN-UHFFFAOYSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- DKPFZGUDAPQIHT-UHFFFAOYSA-N Butyl acetate Natural products CCCCOC(C)=O DKPFZGUDAPQIHT-UHFFFAOYSA-N 0.000 description 1
- DNVPQKQSNYMLRS-NXVQYWJNSA-N Ergosterol Natural products CC(C)[C@@H](C)C=C[C@H](C)[C@H]1CC[C@H]2C3=CC=C4C[C@@H](O)CC[C@]4(C)[C@@H]3CC[C@]12C DNVPQKQSNYMLRS-NXVQYWJNSA-N 0.000 description 1
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 1
- 229930003316 Vitamin D Natural products 0.000 description 1
- MECHNRXZTMCUDQ-UHFFFAOYSA-N Vitamin D2 Natural products C1CCC2(C)C(C(C)C=CC(C)C(C)C)CCC2C1=CC=C1CC(O)CCC1=C MECHNRXZTMCUDQ-UHFFFAOYSA-N 0.000 description 1
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000002050 anti-rickettsial effect Effects 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 239000012156 elution solvent Substances 0.000 description 1
- 229960002061 ergocalciferol Drugs 0.000 description 1
- DNVPQKQSNYMLRS-APGDWVJJSA-N ergosterol Chemical compound C1[C@@H](O)CC[C@]2(C)[C@@H](CC[C@@]3([C@@H]([C@H](C)/C=C/[C@H](C)C(C)C)CC[C@H]33)C)C3=CC=C21 DNVPQKQSNYMLRS-APGDWVJJSA-N 0.000 description 1
- 230000001815 facial effect Effects 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000003791 organic solvent mixture Substances 0.000 description 1
- 238000005192 partition Methods 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 235000019166 vitamin D Nutrition 0.000 description 1
- 239000011710 vitamin D Substances 0.000 description 1
- 150000003710 vitamin D derivatives Chemical class 0.000 description 1
- 235000001892 vitamin D2 Nutrition 0.000 description 1
- 239000011653 vitamin D2 Substances 0.000 description 1
- MECHNRXZTMCUDQ-RKHKHRCZSA-N vitamin D2 Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)/C=C/[C@H](C)C(C)C)=C\C=C1\C[C@@H](O)CCC1=C MECHNRXZTMCUDQ-RKHKHRCZSA-N 0.000 description 1
- 229940046008 vitamin d Drugs 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Steroid Compounds (AREA)
Abstract
Description
【発明の詳細な説明】
〈産業上の利用分野〉
本発明はハラタケ属（Ａｇａｒｉｃｕｓ）のキノコであ
るカワリハラタケ（通称ヒメマツタメケ）（Ａｇａｒｉ
ｃｕｓｂｌａｚｅｉ）の子実体中に存在する新規なエル
ゴステロール誘導体（１）及びエルゴステロール誘導体
（■、ＩＩＩ）の製法に関する。更に詳細には、癌細胞
に対して優れた薬理効果を有するエルゴステロール誘導
体（１、ＩＩ 、Ｉ）に関する。[Detailed Description of the Invention] <Industrial Application Field> The present invention uses Agaricus mushrooms (commonly known as Himematsutameke), which are mushrooms of the genus Agaricus.
The present invention relates to a method for producing a novel ergosterol derivative (1) and a novel ergosterol derivative (■, III) present in the fruiting body of A. cusblazei. More specifically, the present invention relates to ergosterol derivatives (1, II, I) that have excellent pharmacological effects on cancer cells.
〈従来の技術〉
エルゴステロールはキノコに多く含まれ、紫外線照射に
より、エルゴカルシフェロール（ビタミンＤ、）となり
、抗くる病因子として利用されている。またサルノコシ
カケ科のキノコ（ハリＪ１見雇リーｖｅｒｓｉｃｏｌｏ
ｒ）から分離されたエルゴステロール誘導体３β、５α
、６β、９α−トリヒドロキシエルゴスタ−７，２２−
ジエン−６−オン（ＩＩ［）と３β、５α、９α−トリ
ヒドロキシ−６β−メトキシエルゴスタ−７，２２−ジ
エンは肝臓癌細胞に対して殺細胞効果を有し、ｒＴｅｔ
ｒａｈｅｄｒｏｎＪ 、 ３９．２７７９〜２７８５
（１９８３）に記載されている。<Prior Art> Ergosterol is contained in large amounts in mushrooms, and upon irradiation with ultraviolet rays, it becomes ergocalciferol (vitamin D), which is used as an anti-rickets factor. Also, mushrooms of the family Salmonaceae (Hari J1 Mikarei versicolo)
Ergosterol derivatives 3β, 5α isolated from r)
, 6β,9α-trihydroxyergosta-7,22-
Dien-6-one (II[) and 3β,5α,9α-trihydroxy-6β-methoxyergosta-7,22-diene have cytocidal effects on liver cancer cells, and rTet
rahedronJ, 39.2779-2785
(1983).
〈発明が解決しようとしている問題点〉本発明の目的は
、前記従来の技術のエルゴステロール誘導体とは構造を
異にする、新規でしかも優れた殺細胞作用を有するエル
ゴステロール誘導体を提供することと、特許請求範囲第
２項に示されるエルゴステロール誘導体の効率のよい製
法を提供することにある。<Problems to be Solved by the Invention> The purpose of the present invention is to provide a novel ergosterol derivative having an excellent cell-killing effect, which has a different structure from the conventional ergosterol derivatives. The object of the present invention is to provide an efficient method for producing ergosterol derivatives as set forth in claim 2.
〈問題点を解決するための手段〉
本発明者らはキノコより新規な抗癌剤を見出すべく種々
の研究を続けた結果、カワリハラタケから抽出された物
質がヒト子宮頚癌細胞（ＨｅＬａ Ｓｓ）に対して、殺
細胞効果を有することを知った。そこで、さらに研究を
続け、上記殺細胞性を有する物質が新規化合物であり、
前記式（１）で示される３β、５α−ジヒドロキシ−６
β−メトキシエルゴスタ−７，２２−ジエンであること
を確認した。また同時に、ヒメマツタケより２種のエル
ゴステロール誘導体ＣＵ ）（ｍ ）を効率良く分離す
ること１こ成功し、本発明の完成に至った。<Means for Solving the Problems> The present inventors have continued various studies to find novel anticancer drugs from mushrooms, and have found that substances extracted from Kawariharatake have a positive effect on human cervical cancer cells (HeLa Ss). , was found to have a cell-killing effect. Therefore, we continued our research and found that the above-mentioned cell-killing substance was a new compound.
3β,5α-dihydroxy-6 represented by the above formula (1)
It was confirmed that it was β-methoxyergosta-7,22-diene. At the same time, we succeeded in efficiently separating two types of ergosterol derivatives CU ) (m ) from Hiemematsutake, leading to the completion of the present invention.
本発明の化合物は、カワリハラタケの子実体より有機溶
媒で抽出される。有機溶媒の例としては、メタノール、
エタノール、アセトン、酢酸エチル、クロロホルム、等
が挙げられる。抽出は通常室温で行われるが、必要に応
じて加熱還流することもできる。抽出時間は通常１〜７
２時間である。このようにして得られた抽出物は濾過又
は遠心分離あるいはこれらを組み合わせることにより固
液分離される。分離された抽出物は多量の不純物が含ま
れるので、この抽出液から該物質を分離するには水・有
機溶媒混合系で液・液分配抽出することが好ましい。上
記有機溶媒の例としては酢酸エチル、クロロホルム、ジ
エチルエーテル等が挙げられる。The compound of the present invention is extracted from the fruiting body of Kawariharatake with an organic solvent. Examples of organic solvents include methanol,
Examples include ethanol, acetone, ethyl acetate, chloroform, and the like. Extraction is usually carried out at room temperature, but can also be heated to reflux if necessary. Extraction time is usually 1-7
It is 2 hours. The extract thus obtained is subjected to solid-liquid separation by filtration, centrifugation, or a combination thereof. Since the separated extract contains a large amount of impurities, it is preferable to perform liquid-liquid partition extraction using a mixed water/organic solvent system in order to separate the substance from the extract. Examples of the organic solvent include ethyl acetate, chloroform, diethyl ether, and the like.
該物質は、水に不溶で、上記有機溶媒に易溶でかつ中性
物質であることから、上記の如く得られた抽出物を塩基
性、酸性条件下で一回又は繰り返し水・有機溶媒、液・
液分配抽出を行うことにより分離精製を行うことができ
る。即ち、上記、有機溶媒混合系にアンモニア、炭酸ナ
トリウム、重炭酸ナトリウム、水酸化ナトリウム等を加
え酸性物質を水層に転溶して除いた後、有機溶媒層を回
収し、次に酸たとえば酢酸、塩酸等の水溶液を加えて塩
基性物質を水層に転溶して除く。これらの操作を一回又
は繰り返し行うことにより不要成分を取り除くことがで
きる。Since the substance is insoluble in water, readily soluble in the organic solvent, and neutral, the extract obtained as described above is treated once or repeatedly with water/organic solvent, under basic or acidic conditions. liquid·
Separation and purification can be performed by performing liquid partition extraction. That is, after adding ammonia, sodium carbonate, sodium bicarbonate, sodium hydroxide, etc. to the above organic solvent mixture system and removing the acidic substances by dissolving them in the aqueous layer, the organic solvent layer is collected, and then an acid such as acetic acid is added. , add an aqueous solution such as hydrochloric acid to remove the basic substance by dissolving it in the aqueous layer. Unwanted components can be removed by performing these operations once or repeatedly.
さらに精製を進めるために得られた抽出物は続いて吸着
クロマトグラフィーに付される。吸着クロマトグラフィ
ーには例えば、シリカゲルカラムクロマトグラフィー又
は薄層クロマトグラフィーが用いられる。展開溶媒とし
ては例えばベンゼン、クロロホルム、ジクロロメタン、
酢酸エチル、酢酸ブチル、四塩化炭素、メタノール、エ
タノール等の１つ又はそれらの混合物が適宜選択される
。The obtained extract is subsequently subjected to adsorption chromatography for further purification. For example, silica gel column chromatography or thin layer chromatography is used for adsorption chromatography. Examples of developing solvents include benzene, chloroform, dichloromethane,
One or a mixture thereof such as ethyl acetate, butyl acetate, carbon tetrachloride, methanol, and ethanol is selected as appropriate.
さらに精製を進めるためにセファデックスＬＨ−２０（
ファルマシア社製）等のゲル濾過剤を用いることも育効
である。Sephadex LH-20 (
It is also effective to use gel filtration agents such as those manufactured by Pharmacia.
このようにして本発明の新規化合物を含む３種の殺細胞
活性エルゴステロール誘導体を分離することができる。In this way, three cell-killing active ergosterol derivatives, including the novel compounds of the present invention, can be separated.
以下に、本発明の新規化合物（Ｉ）の物理化学的性質を
述べる。The physicochemical properties of the novel compound (I) of the present invention will be described below.
（１）分子量 ＝４４４
（２）比施光度 ： 〔α） Ｌ” ＝ ６０．
８゜（３）赤外線吸収スペクトル ： ３４００．１
６４０（４）溶剤に対する溶解性 ： 酢酸エチル、ク
ロロホルムに可溶、メタノール、エタノールにやや可溶
、水に不溶
（５）呈色反応 二 ローゼンハイム反応陽性（６）塩
基性、中性、酸性の区別 ： 中性物質（７）物質の色
及び形状 ： 無色、シロップ状以上、本発明の新規化
合物は３β、５α−ジヒドロキシ−６β−メトキンエル
ゴスタ−７，２２−ジエンであることが本発明者等によ
って決定された。(1) Molecular weight = 444 (2) Specific light intensity: [α) L” = 60.
8゜(3) Infrared absorption spectrum: 3400.1
640 (4) Solubility in solvents: Soluble in ethyl acetate and chloroform, slightly soluble in methanol and ethanol, insoluble in water (5) Color reaction 2 Rosenheim reaction positive (6) Basic, neutral, acidic Distinction: Neutral Substance (7) Color and Shape of Substance: Colorless, syrup-like or above, the novel compound of the present invention is 3β,5α-dihydroxy-6β-methquinergosta-7,22-diene. It was determined by the following persons.
本発明によって得られる新規化合物を含む３種のエルゴ
ステロール誘導体は次の試験例に示すように癌細胞に対
し顕著な殺細胞作用を有する。Three types of ergosterol derivatives, including the novel compound obtained by the present invention, have a remarkable cytocidal effect on cancer cells, as shown in the following test examples.
〈試験例〉
ヒト子宮頚癌由来の培養株細胞（ＨｅＬａ Ｓ３）に対
するｉｎ ｖｉｔｒｏ殺細胞作用
プラスチック製９６穴マイクロプレート（Ｃｏｒｎｉｎ
ｇ社製）に２ Ｘ １０’Ｃｅ１ｌｓ／ｍ（７となるよ
うに牛胎児血清１０％を含むＥａｇｌｅ’ ｓ Ｍ Ｅ
Ｍ培地に浮遊懸濁させたＨｅＬａ５ｓ細胞を１穴当た
り０．２ｍｆ２播種した。これを５％炭酸ガス培養器中
で３７℃、２４時間培養後、この培養液中に薬剤溶液を
５μＱ加え、さらに上記と同様の条件下で７２時間培養
し、培地上清を除いた上で細胞をメタノールで固定化し
、ギムザ染色後、細胞増殖の状態を鏡検した。薬剤とし
ては、本発明の新規化合物を含む３種のエルゴステロー
ル誘導体を種々の濃度となるようにメタノールに溶解し
、作製した。<Test Example> In vitro cell killing effect on cultured cell line derived from human cervical cancer (HeLa S3) Plastic 96-well microplate (Cornin
Eagle's ME containing 10% fetal bovine serum to give 2 x 10'Cells/m (7)
HeLa5s cells suspended in M medium were seeded at 0.2 mf2 per well. After culturing this in a 5% carbon dioxide incubator at 37°C for 24 hours, 5 μQ of the drug solution was added to this culture solution, and further cultured for 72 hours under the same conditions as above, and after removing the medium supernatant. Cells were fixed with methanol, stained with Giemsa, and then microscopically examined for cell proliferation. Drugs were prepared by dissolving three types of ergosterol derivatives, including the novel compound of the present invention, in methanol at various concentrations.
面述の鏡検下で、殺細胞効果を調べ、生細胞数が、全く
認められない培地中の薬剤の最小濃度を最終有効濃度と
した。その結果、第１表に示す通り、本発明の新規化合
物を含むエルゴステロール誘導体に著しい殺細胞作用が
認められた。The cell-killing effect was examined under facial microscopic examination, and the lowest concentration of the drug in the medium at which no viable cell count was observed was determined as the final effective concentration. As a result, as shown in Table 1, the ergosterol derivatives containing the novel compound of the present invention were found to have a significant cell-killing effect.
〈実施例〉
以下、本発明の実施例を示すが、これによって限定され
るものではない。<Examples> Examples of the present invention will be shown below, but the present invention is not limited thereto.
カワリハラタケ子実体５Ｋｇにアセトン１０（を加えホ
モジナイズし抽出を行った。この抽出物を濾過し、が液
を減圧下で濃縮した。さらに濃縮物を１１２の酢酸エチ
ルで３回液・液分配抽出した。得られた酢酸エチル層を
飽和炭酸水素ナトリウム水溶液で洗浄し、次いでＩＮ塩
酸で洗浄した。このようにして得られた酢酸エチル層を
濃縮し、抽出物１４．９ｇを得た。この抽出物をクロロ
ホルム：メタノール（９４： ６）に溶解し、Ｗａｋｏ
ゲルＣ−２００，４５０ｇを用いてカラムクロマトグラ
フィーを行った。この時の溶出溶媒としてはクロロホル
ム：メタノール（９４：６）を用いた。次に薄層クロマ
トグラフィーによって繰り返し精製することにより、新
規エルゴステロール誘導体３β、５α−ジヒドロキシ−
６β−メトキシエルゴスタ−７，２２−ジエン（Ｉ ）
３．４ｍｇと既知のエルゴステロール誘導体として、３
β、５α、６β、９α−テトラヒドロキシエルゴスタ−
７，２２−ジエン（［［）６．３Ｂ、モして ３β、５
α、９α−トリヒドロキシエルゴスタ−７，２２−ジエ
ン−６−オン＜ｍ ）５．７ｍｇを得た。Acetone (10 kg) was added to 5 kg of Kawariharatake fruiting body and homogenized to perform extraction. The extract was filtered, and the liquid was concentrated under reduced pressure. The concentrate was then subjected to liquid-liquid partition extraction three times with 112 ethyl acetate. The obtained ethyl acetate layer was washed with a saturated aqueous sodium bicarbonate solution, and then washed with IN hydrochloric acid.The ethyl acetate layer thus obtained was concentrated to obtain 14.9 g of an extract. was dissolved in chloroform:methanol (94:6), and Wako
Column chromatography was performed using 450 g of Gel C-200. Chloroform:methanol (94:6) was used as the elution solvent at this time. Then, by repeated purification by thin layer chromatography, a new ergosterol derivative 3β,5α-dihydroxy-
6β-methoxyergosta-7,22-diene (I)
As an ergosterol derivative known as 3.4 mg, 3
β, 5α, 6β, 9α-tetrahydroxyergoster
7,22-diene ([[)6.3B, 3β, 5
5.7 mg of α,9α-trihydroxyergosta-7,22-dien-6-one <m ) was obtained.
Claims (2)
エルゴスタ−７，２２−ジエン（ I ）である化合物。(1) A compound that is 3β,5α-dihydroxy-6β-methoxyergosta-7,22-diene (I) represented by the structural formula (I) ▲Mathematical formula, chemical formula, table, etc.▼(I).
示される３β，５α，６β，９α−テトラヒドロキシエ
ルゴスタ−７，２２−ジエン（II）または３β，５α，
９α−トリヒドロキシエルゴスタ−７，２２−ジエン−
６−オン（III）である化合物の製法。(2) Structural formula ▲ Numerical formula, chemical formula, table, etc. ▼ R_1=-OH...(II) R_2==O...(III) [In the formula, R_1 represents a hydroxyl group or a ketone group. ] 3β,5α,6β,9α-tetrahydroxyergosta-7,22-diene (II) or 3β,5α,
9α-trihydroxyergosta-7,22-diene-
A method for producing a compound that is 6-one (III).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP7187988A JPH01246299A (en) | 1988-03-28 | 1988-03-28 | Ergosterol derivative and production thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP7187988A JPH01246299A (en) | 1988-03-28 | 1988-03-28 | Ergosterol derivative and production thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH01246299A true JPH01246299A (en) | 1989-10-02 |
Family
ID=13473245
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP7187988A Pending JPH01246299A (en) | 1988-03-28 | 1988-03-28 | Ergosterol derivative and production thereof |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH01246299A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100559263B1 (en) * | 2004-08-31 | 2006-03-15 | 한국생명공학연구원 | A method of preparing ergosterol epoxide and physiologically acceptable salts thereof from Lentinus edodes |
| JP2008115158A (en) * | 2006-10-13 | 2008-05-22 | Hiroko Ito | Steroid derivative, method for producing the same and apoptosis-inducing agent |
| RU2477635C1 (en) * | 2012-02-14 | 2013-03-20 | Глеб Иванович Кирьянов | Method of producing incisterol |
| WO2013073085A1 (en) * | 2011-11-19 | 2013-05-23 | ホクト株式会社 | Apoptosis inducer |
| CN105338957A (en) * | 2013-06-24 | 2016-02-17 | 株式会社爱茉莉太平洋 | External use skin composition, containing lentinula edodes-derived ergosterol |
-
1988
- 1988-03-28 JP JP7187988A patent/JPH01246299A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100559263B1 (en) * | 2004-08-31 | 2006-03-15 | 한국생명공학연구원 | A method of preparing ergosterol epoxide and physiologically acceptable salts thereof from Lentinus edodes |
| JP2008115158A (en) * | 2006-10-13 | 2008-05-22 | Hiroko Ito | Steroid derivative, method for producing the same and apoptosis-inducing agent |
| WO2013073085A1 (en) * | 2011-11-19 | 2013-05-23 | ホクト株式会社 | Apoptosis inducer |
| JPWO2013073085A1 (en) * | 2011-11-19 | 2015-04-02 | ホクト株式会社 | Apoptosis inducer |
| RU2477635C1 (en) * | 2012-02-14 | 2013-03-20 | Глеб Иванович Кирьянов | Method of producing incisterol |
| CN105338957A (en) * | 2013-06-24 | 2016-02-17 | 株式会社爱茉莉太平洋 | External use skin composition, containing lentinula edodes-derived ergosterol |
| CN105338957B (en) * | 2013-06-24 | 2018-08-17 | 株式会社爱茉莉太平洋 | Include the Dermatologic preparation composition of the ergosterol from white flower mushroom |
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